Expression of Kita-Kyushu Lung Cancer Antigen-1 as Detected by a Novel Monoclonal Antibody in Gastric Cancer.

BACKGROUND/AIM: Kita-Kyushu lung cancer antigen-1 (KK-LC-1) is a known cancer/testis antigen. Our group has previously shown KK-LC-1 gene expression in gastric cancer. However, could not be detected the KK-LC-1 protein due to the lack of an appropriate antibody. Here, we assessed our original monoclonal antibody (Kmab34B3) and, using it, assessed the expression of KK-LC-1 in gastric cancer. PATIENTS AND METHODS: We evaluated an original monoclonal antibody against KK-LC-1 (Kmab34B3), and used this antibody to compare KK-LC-1 protein expression in tumour and non-tumour stomach cells from gastric cancer patients. RESULTS: Kmab34B3 stained testicular germ cells, and tumour cells in nine out of 11 (82%) specimens. In non-tumorous areas, Kmab34B3 stained 13 out of 29 (45%) pyloric gland specimens. Furthermore, Kmab34B3 also stained intestinal metaplasia positive and negative areas. CONCLUSION: Kmab34B3 was able to detect KK-LC-1 protein within tumour cells and the pyloric gland where the gene has been shown to be expressed. Therefore, it might be an attractive tool for detecting KK-LC-1 expression in precancerous and cancerous stomach cells.

Immunohistochemical identification and localisation of gastrin and somatostatin in endocrine cells of human pyloric gastric mucosa.

The detailed description of the distribution of endocrine cells G and D producing important hormones that regulate activation of other cells in the human stomach may be a valuable source of information for opinions about mucosa changes in different diseases of the alimentary tract. The density and distribution of immunoreactive G and D cells in the pylorus of humans (donors of organs) were evaluated. The pylorus samples were collected after other organs were harvested for transplantation. The number of G cells in the pyloric mucosa of healthy people was higher than the number of D cells. G and D cells were distributed between columnar cells of epithelium mucosa. Multiform endocrine cells generally occurred: gastrin in the middle third of the mucosa and somatostatin cells in the basal half of the pyloric mucosa. The investigation of the pyloric part of the healthy human stomach showed a characteristic distribution of cells that reacted with antisera against gastrin and somatostatin.

Pyloric tonsil as a novel gut-associated lymphoepithelial organ of the chicken.

The pyloric tonsil is a novel peripheral lymphoepithelial organ of the gastrointestinal tract in the chicken. It forms a complete lymphoid ring at the beginning of the duodenum, where crypts of Lieberkühn are transformed to tonsillar crypts with lymphoepithelial lining. The oesophageal (described previously) and pyloric tonsils are characteristic of the chicken, while they are absent in mammals. The lymphoid system develops from the middle germ layer, the mesoderm, and forms connections with the ecto- and endoderm, namely the skin and gut, respectively. These connections are based on the lymphoepithelial lining of the crypts, and provide gates for environmental antigens. Recent findings, taken together with the literature, suggest that in birds the lymphoid system forms connections with the endoderm-derived organs that are anatomically and histologically more extensive than the ectoderm-derived ones, which may be explained by the absence of regional lymph nodes, and the less developed lymphoid circulation of the skin.

Involvement of dendritic cell response to resistance of stomach carcinogenesis caused by N-methyl-N'-nitro-N-nitrosoguanidine in rats.

The involvement of immune response in the resistance of chemically induced stomach cancer was studied in a resistant rat strain (Buffalo) and a sensitive rat strain (ACI). Groups of 10 male Buffalo and ACI rats, 6 weeks of age, were given drinking water with or without N-methyl-N'-nitro-N-nitrosoguanidine (MNNG; 100 mg/l) for 14 days. Total RNA was isolated from the stomach pyloric mucosa from five rats, and cDNA was prepared with reverse transcriptase. Tissue sections of the stomach pyloric mucosa from five rats were stained with antibodies recognizing molecules expressed by various immune cells. Reverse transcription-PCR (RT-PCR), competitive RT-PCR, and Northern blot demonstrated that the expression of MHC class II group genes [MHC class II, MHC class II-associated invariant chain (Ii), CD4 and IgM (B cell marker)], MHC class I group genes (MHC class I and CD8), B7-1 (costimulator on dendritic cells), and CD28 (receptor to B7 on T cells) in the pyloric mucosa was elevated by MNNG in both rat strains but was elevated to a 4-7-fold greater extent in Buffalo rats than in ACI rats. These genes were scarcely expressed in control rats. Histochemical antibody staining after MNNG exposure showed a greater number of cells stained with monoclonal antibody to Ii, OX-62 (dendritic cell marker), and ED-1 (dendritic cell and macrophage common marker) in the interstitial tissue of the pyloric mucosa of Buffalo rats compared with ACI rats. Cell proliferation, as measured by 5-bromo-2-deoxyuridine (BrdUrd)-labeling indices, revealed the presence of BrdUrd-labeled cells only among epithelial cells in the proliferative zone; cells in the interstitial tissue were not labeled with BrdUrd. The results suggest the involvement of dendritic cell response in the resistance to the MNNG induction of stomach carcinogenesis in rats.

The pathology of infantile hypertrophic pyloric stenosis after healing.

INTRODUCTION: Infantile hypertrophic pyloric stenosis (IHPS) is a common surgical affection of unknown etiology. The muscular hypertrophy is known to resolve within a few months after pyloromyotomy (PM). The pathology of IHPS has been studied extensively at the time of PM, but the fate of the pylorus after healing remains unknown. MATERIALS AND METHODS: We had the rare opportunity to study two pyloric biopsy specimens obtained 4 months and 2 years (respectively) after an uncomplicated PM for IHPS. They were compared with the initial specimen in one case, with 26 other specimens of IHPS, and with five normal controls. Immunohistochemistry using the avidin-biotin complex (ABC) system was performed for S-100 and nerve growth factor receptor, as markers for the enteric nervous system, and for the tyrosine kinase receptor c-kit, as a marker for the interstitial cells of Cajal (pacemaker cells). NADPH-diaphorase histochemistry was performed as a marker for the neuronal enzyme nitric oxide synthase, which produces the inhibitory neurotransmitter nitric oxide. RESULTS: In both cases of IHPS, after healing, the circular musculature was not hypertrophic. For all markers studied, the distribution appeared similar to that in the normal pylorus. In contrast, all specimens obtained at the time of PM displayed a severe reduction of the different markers in the hypertrophic musculature. DISCUSSION: The pathological features observed in the circular layer in IHPS appear to resolve within a few months after PM. This suggests that the involvement of the enteric nervous system in IHPS might be milder than generally assumed. The etiology remains obscure, but our occasional observations may provide new insight into the pathophysiology of IHPS, and are in agreement with the excellent longterm clinical outcome for IHPS.

Cellular differentiation and development of pyloric mucosal metaplasia in the human gall-bladder.

The development of pyloric mucosal metaplasia (PMM) with regard to cellular differentiation in the human gallbladder was studied by mucin staining (paradoxical concanavalin A (Con A), galactose oxidase-Schiff (GOS) and alcian blue (pH 2.5) PAS (AB-PAS) and immunohistochemistry (pepsinogen II (PgII) and SH-9, and proliferating cell nuclear antigen (PCNA). PMM was divided into three stages of development by three-dimensional (3D) computer graphic reconstruction analysis. In the early stage, a transitional zone of PCNA positive cells was observed between areas of SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells in flat monolayered epithelium. In the middle stage, shallow pits became apparent as areas enlarged, with these becoming deeper in the advanced stage, whereby SH-9 and/or GOS reactive cells and class III and/or Pg II positive cells were observed at the upper and lower portion of the pits, respectively, with PCNA-positive cells forming a narrow zone between the two cell populations. Consequently, the structure of PMM gradually resembles that of the normal gastric pyloric mucosa.

[Cellular and humoral immunity in duodenal peptic ulcer with gastrin cell hyperplasia]. / Kletochnyi i gumoral'nyi immunitet pri iazvennoi bolezni dvenadtsatiperstnoi kishki s giperplaziei gastrinovykh kletok.

A total of 61 patients with frequently relapsing duodenal ulcer were examined. Of these, 18 patients were in the acute phase and 43 experienced remission. Using 1 mm2 of the mucosa, measurements were made of the counts of duodenal and pyloric G-cells (by immunomorphologic assay), of the absolute and relative counts of T and B lymphocytes, the content of IgA, IgM and IgG, histamine and serotonin (by fluorometry) in the blood, and of the concentration of uropepsin in the urine. In the stages of exacerbation and remission, the patients suffering from duodenal ulcer with hyperplasia of G-cells manifested, as compared with the analogous patients without hyperplasia, a decrease of the absolute and relative counts of T cells, especially of those of B cells, combined with a rise of the content of IgM and IgG during exacerbation, followed by its returning to normal in the phase of remission. Over one year part of the duodenal ulcer patients with hyperplasia of G-cells received preventive treatment with ranitidine, which resulted in a tendency towards the lowering of the count of pyloric G-cells and the rise of the absolute and relative counts of T cells.

Relationship between Campylobacter pylori and gastritis in healthy humans after administration of placebo or indomethacin.

Endoscopic and microscopic appearances of antral and fundic mucosa were correlated with the presence or absence of Campylobacter pylori--and with plasma immunoglobulin G antibodies to that organism--in 23 healthy volunteers, 12 of whom had received indomethacin and 11 of whom had received no medication. Antral C. pylori, found in 9 of 23 biopsy specimens (3 of 11 controls, 6 of 12 indomethacin-treated patients; not significantly different), correlated strongly with histologic evidence of active superficial antral gastritis (p less than 0.005), but not with the endoscopic appearance of the antrum. In contrast to the antrum, fundic C. pylori, found in 14 of 23 biopsy specimens (61%), were frequently associated with histologically and endoscopically normal fundic mucosa. Campylobacter pylori-associated active antral gastritis occurred only in subjects whose fundus harbored this organism. Plasma immunoglobulin G antibody titers to C. pylori were highest in subjects with Campylobacter-associated antral gastritis and lowest in subjects without gastric Campylobacter. These studies suggest that healthy humans may harbor C. pylori in their proximal stomach without apparent ill effects. In some of these individuals, the organism also involves the antrum and is associated with active gastritis.

Immunohistologic pattern of type 1 (Lea, Leb) and type 2 (X, Y, H) blood group-related antigens in the human pyloric and duodenal mucosae.

Type 1 antigens (Lea and Leb) of the pyloric mucosa are restricted to the mucus cells of the surface epithelium and the neck. In the duodenum they are present in absorptive and goblet cells of duodenal villi as well as in the glands of Lieberkühn. Type 2 antigens (X, Y, and H) are also present in these areas of the mucosa, and both type 1 and type 2 antigens, in this superficial location, are under the control of the secretor gene. In addition, type 2 antigens are also present in the mucus cells of pyloric and Brunner's glands, but in this deep location they are not controlled by the secretor gene. These results suggest the existence of two differentiation patterns for the expression of glycosyltransferases in the stem cells. The upward differentiation process would favor the expression of the alpha-2-L-fucosyltransferase coded by the Se gene, and the downward differentiation process would favor the expression of the alpha-2-L-fucosyltransferase coded by the H gene. The knowledge of the distribution and the genetic control of type 1 and type 2 antigenic determinants in normal gastrointestinal tract may help to interpret the modifications in the expression of these antigens in pathologic conditions such as neoplasia.

Immunohistochemical studies on the development of prochymosin- and pepsinogen-containing cells in bovine abomasal mucosa.

1. Prochymosin- and pepsinogen-containing cells in abomasal mucosa were simultaneously localized by means of an indirect immunofluorescence technique with specific rabbit antibodies. 2. In the young milk-fed calves all the chief cells and several mucous neck cells produced both prochymosin and pepsinogen. 3. By elution of the first antibody and restaining of the same section it was demonstrated that the same cells were producing both prochymosin and pepsinogen. 4. In concentrate-fed calves and older cattle all chief cells and almost all mucous neck cells produced pepsinogen, while prochymosin was produced only from the chief cells situated in the upper part of the base in the gastric gland. 5. The number of prochymosin-containing cells was highly correlated to the milk-feeding of the animal. 6. The present data suggest that the non-differentiated cell in the neck region is initially capable of producing both prochymosin and pepsinogen during the proliferation phase to a mature chief cell, via mucous neck cell. The ability to produce prochymosin is however only retained if the mucosa is stimulated by milk-feeding. Dietary factors may thus be of importance for the differentiation of the gastrointestinal epithelium.

[Interepithelial lymphocytes of the gastric mucosa in peptic ulcer]. / Mezhépitelial-nye limfotsity slizistoí obolochki zheludka pri iazvennoí bolezni.

Unlike intestinal interepithelial lymphocytes (IEL), gastric IEL have not been studied. Gastric mucosa from 27 stomachs resected for peptic ulcer was examined. The intact high prismatic superficial epithelium contained 51.38 +/- 4.39 IEL per 1000 epitheliocytes, foveolar epithelium 56.31 +/- 5.77. In chronic gastritis, in both superficial and foveolar epithelium the number of IEL was increased nearly 3-fold and was 161.3 +/- 21.08 and 154.81 +/- 15.03, respectively. In these cases the number of lymphocytes in lamina propria did not increase. The increase in the number of IEL correlated with a marked increase of plasma cells in lamina propria and with rapid epithelial proliferation. The epithelium lining of the stomach as well as that of the small intestine is the site of differentiation of the immunological activity of lymphoid cells as indicated by mitosis figures in IEL as well as the presence in superficial epithelium of plasma cells the appearance of which may be explained by disorders in differentiation of B-lymphocytes and maturation of plasma cells in the epithelial sheet. Infiltration of proliferating epithelium with IEL suggests that they take part in stimulation of cell renewal in gastric mucosa.

Immunoreactivities of gastrin (G-) cells. I. dilution-dependent staining of G-cells by antisera and non-immune sera.

Results of immunocytochemical studies reported by several laboratories suggest that gastrin (G-) cells of the stomach show immunoreactivities for various pituitary hormones (ACTH, met-enkephalin, beta-endorphin and growth hormone) in addition to gastrin. By reinvestigating the immunocytochemistry of G-cells we found that these cells exhibited reactivities towards a variety of antisera against enteric, pancreatic and hypophyseal hormones. Gastrin cells can also be "immunostained" by antisera towards proteins unrelated to any peptide hormones (e.g. alpha-fetoprotein antiserum) and by nonimmune sera. Thus the specificity of immunocytochemical findings in G-cells seems to be uncertain. According to our findings the polyvalent immunoreactivities of G-cells may be caused by a distinct binding capacity for IgG molecules. This binding of IgG to G-cells seems to be mediated by the Fab fragments of the IgG molecules which may behave like a basic dye and therefore "immunostain" anionic components within G-cells. Thus the significance of the immunocytochemical proof of peptide hormones within G-cells is limited unless extended specificity controls have been performed. The results of specificity controls performed in this study (adsorption controls, use of ascending dilutions of the primary and secondary antisera, comparison of crude antisera and affinity chromatographically purified antibodies) suggest that corticotropin-lipotropin related peptides are not contained in G-cells.

Effect of seasonal variation on lymphoid tissues of the lizards, Mabuya quinquetaeniata Licht. and Uromastyx aegyptia Forsk.

The thymus in the lizards Mabuya quinquetaeniata and Uromastyx aegyptia is highly involuted in winter but exhibits in the other seasons a rich lymphoepithelial organization. The splenic white pulp is severely depleted in winter but is extensively developed in spring, summer and autumn. In these seasons, the splenic lymphoid tissue of Mabuya occurs in a continuous phase throughout the organ obscuring the red pulp, whereas in Uromastyx the white pulp remains localized as periarteriolar aggregates and the red pulp is always clearly delineated. In both lizard species, gut-associated lymphoid tissue is well represented, especially in the large intestines and in Mabuya it is almost similar in different seasons. In Uromastyx, in winter, lymphoid nodules are only found in the caecum and the colon, but during warm seasons, inumerable nodules are distributed throughout the gut. The findings are important for a clearer understanding of immunologic competence in reptiles.

Immunohistochemical demonstration of somatostatin-positive cells in the canine gut.

Somatostatin-immunoreactive cells in the canine gut were investigated by an indirect immunofluorescent technique. Somatostatin-positive cells were demonstrated in the neck portion of the pyloric gland and in the bottom of the intestinal crypt, and these cells were distinguished from those in other organs by the presence of a cytoplasmic process reaching the gut lumen (external environment). Possible functional meaning of this cytoplasmic process was discussed.

Distribution and properties of glucagon immunoreactivity in the digestive tract of various mammals: an immunohistochemical and immunochemical study.

Numerous epithelial cells in the oxyntic gland area of the stomach of cat and dog appear to store pancreatic-type glucagon, as evidenced by immunohistochemistry and--in the case of the dog--also by radioimmunoanalysis. No glucagon immunoreactive cells are found in the antropyloric mucosa. Disseminated cells in the intestinal mucosa store gut-type glucagon. In all species studied, these cells predominate in ileum and colon.